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Introduction

This protocol is intended for magnetic isolation of CD4+CD25+ regulatory T cells from human mononuclear cells (MNCs). The CD4+CD25- cell fraction can be used as effector cells in downstream inhibitory assays. The supplied protocol describes magnetic labeling and isolation from 1 x 108 MNCs. In the first step the non-CD4+ MNC are labelled with Antibody Mix Human CD4. In the second step Depletion MyOne Dynabeads® are added to remove the non-CD4+ MNCs. In the third step Dynabeads® CD25 are added to the CD4+ T cells to capture the CD4+CD25+ T cells and in the last step Dynabeads® CD25 are removed from the cells.
Downstream Applications

Isolated cells may be used directly in any downstream application including flow cytometry and cell expansion protocols.

Additional Materials Required

Materials that are not included, but are needed to perform the entire protocol:

This product is stable until the expiry date stated on the label when stored unopened at 2-8°C. Store opened vials at 2-8°C and avoid bacterial contamination. Keep Dynabeads® in liquid suspension during storage and all handling steps, as drying will result in reduced performance. Resuspend well before use.

Warnings And Limitations

This product is for research use only. Not intended for any animal or human therapeutic or diagnostic use unless otherwise stated. Follow appropriate laboratory guidelines. This product contains 0.02% sodium azide as a preservative, which is cytotoxic.

Avoid pipetting by mouth!

Sodium azide may react with lead and copper plumbing to form highly explosive metal azides. When disposing through plumbing drains, flush with large volumes of water to prevent azide build up. Certificate of Analysis (CoA) is available upon request. Material Safety Data Sheet (MSDS) is available at .